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. 2011 Dec 1;287(4):2836–2842. doi: 10.1074/jbc.M111.300749

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© 2012 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 2. — A crucial role of FKP12 for rapamycin inactivation of TOR. A, FKP12 requirement for inhibition of Thr-449 phosphorylation by rapamycin (Rap). WT or fkp12 mutant protoplasts expressing FLAG-tagged S6K1 were treated with rapamycin. Total proteins were analyzed by protein blotting probed with anti-Thr(P)-389 (P-T449) or anti-FLAG (S6K1) antibody. B, RT-PCR analysis of FKP12 expression. C, FKP12 requirement for inhibition of seedling growth by rapamycin. Rapamycin was added at the time of germination for 9 days. D, overexpression of Arabidopsis or human FKP12 enhancing the rapamycin inhibition effect on Thr-449 phosphorylation in leaf cells. FLAG-tagged S6K1 was co-expressed with HA-tagged FKP12s or GFP in protoplasts and treated with rapamycin for 30 min. E, overexpression of Arabidopsis or human FKP12 enhancing rapamycin inhibition effect on seedling growth. Rapamycin was added at the time of germination for 9 days. Scale bar, 5 mm.